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Biol Proced Online. 2011 Jan 31;13:3. doi: 10.1186/1480-9222-13-3.

A quantitative PCR method for measuring absolute telomere length.

Author information

  • 1CSIRO Food and Nutritional Sciences, Nutritional Genomics Laboratory, PO Box 10041, Adelaide BC, SA, 5000, Australia.

Abstract

We describe a simple and reproducible method to measure absolute telomere length (aTL) using quantitative real-time polymerase chain reaction (qPCR). This method is based on the Cawthon method for relative measurement of telomere length (TL) but modified by introducing an oligomer standard to measure aTL. The method describes the oligomer standards, the generation of the standard curve and the calculations required to calculate aTL from the qPCR data. The necessary controls and performance characteristics of the assay are described in detail and compared relative to other methods for measuring TL. Typical results for this assay for a variety of human tissue samples are provided as well as a troubleshooting schedule. This method allows high throughput measurement of aTL using small amounts of DNA making it amenable for molecular epidemiological studies. Compared to the traditional relative TL qPCR assays, the aTL method described in this protocol enables a more direct comparison of results between experiments within and between laboratories.

PMID:
21369534
[PubMed]
PMCID:
PMC3047434
Free PMC Article

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